Intracellular trafficking and ubiquitination of the Schizosaccharomyces pombe amino acid permease Aat1p

Abstract The prp (pre-mRNA processing) mutants of the fission yeast Schizosaccharomyces pombe have a defect in pre-mRNA splicing and accumulate mRNA precursors at a restrictive temperature. One of the prp mutants, prp1-4, also has a defect in poly(A)+ RNA transport. The prp1 + gene encodes a protein of 906 amino acid residues that contains 19 repeats of 34 amino acids termed tetratrico peptide repeat (TPR) motifs, which were proposed to mediate protein-protein interactions. The amino acid sequence of Prplp shares 29.6% identity and 50.6% similarity with that of the PRP6 protein of Saccharomyces cerevisiae, which is a component of the U4/U6 snRNP required for spliceosome assembly. No functional complementation was observed between S. pombe prp1 + and S. cerevisiae PRP6. We examined synthetic lethality of prp1-4 with the other known prp mutations in S. pombe. The results suggest that Prp1p interacts either physically or functionally with Prp4p, Prp6p and Prp13p. Interestingly, the prp1 + gene was found to be identical with the zer1 + gene that functions in cell cycle control. These results suggest that Prp1p/Zer1p is either directly or indirectly involved in cell cycle progression and/or poly(A)+ RNA nuclear export, in addition to pre-mRNA splicing.

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The amino acid permease AAP8 is important for early seed development in Arabidopsis thaliana

Planta ◽

10.1007/s00425-007-0527-x ◽

2007 ◽

Vol 226 (4) ◽

pp. 805-813 ◽

Cited By ~ 73

Author(s):

Roberto Schmidt ◽

Harald Stransky ◽

Wolfgang Koch

Keyword(s):

Arabidopsis Thaliana ◽

Amino Acid ◽

Seed Development ◽

Amino Acid Permease ◽

Early Seed Development

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Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe.

Molecular and Cellular Biology ◽

10.1128/mcb.13.10.6393 ◽

1993 ◽

Vol 13 (10) ◽

pp. 6393-6402 ◽

Cited By ~ 32

Author(s):

M A MacInnes ◽

J A Dickson ◽

R R Hernandez ◽

D Learmonth ◽

G Y Lin ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Schizosaccharomyces Pombe ◽

Excision Repair ◽

Cdna Clones ◽

Recombination Mechanism ◽

Helix Loop Helix ◽

Partial Length ◽

Human Genes ◽

Single Clone

Several human genes related to DNA excision repair (ER) have been isolated via ER cross-species complementation (ERCC) of UV-sensitive CHO cells. We have now isolated and characterized cDNAs for the human ERCC5 gene that complement CHO UV135 cells. The ERCC5 mRNA size is about 4.6 kb. Our available cDNA clones are partial length, and no single clone was active for UV135 complementation. When cDNAs were mixed pairwise with a cosmid clone containing an overlapping 5'-end segment of the ERCC5 gene, DNA transfer produced UV-resistant colonies with 60 to 95% correction of UV resistance relative to either a genomic ERCC5 DNA transformant or the CHO AA8 progenitor cells. cDNA-cosmid transformants regained intermediate levels (20 to 45%) of ER-dependent reactivation of a UV-damaged pSVCATgpt reporter plasmid. Our evidence strongly implicates an in situ recombination mechanism in cDNA-cosmid complementation for ER. The complete deduced amino acid sequence of ERCC5 was reconstructed from several cDNA clones encoding a predicted protein of 1,186 amino acids. The ERCC5 protein has extensive sequence similarities, in bipartite domains A and B, to products of RAD repair genes of two yeasts, Saccharomyces cerevisiae RAD2 and Schizosaccharomyces pombe rad13. Sequence, structural, and functional data taken together indicate that ERCC5 and its relatives are probable functional homologs. A second locus represented by S. cerevisiae YKL510 and S. pombe rad2 genes is structurally distinct from the ERCC5 locus but retains vestigial A and B domain similarities. Our analyses suggest that ERCC5 is a nuclear-localized protein with one or more highly conserved helix-loop-helix segments within domains A and B.

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Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe

Molecular and Cellular Biology ◽

10.1128/mcb.13.10.6393-6402.1993 ◽

1993 ◽

Vol 13 (10) ◽

pp. 6393-6402

Author(s):

M A MacInnes ◽

J A Dickson ◽

R R Hernandez ◽

D Learmonth ◽

G Y Lin ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Schizosaccharomyces Pombe ◽

Excision Repair ◽

Cdna Clones ◽

Recombination Mechanism ◽

Helix Loop Helix ◽

Partial Length ◽

Human Genes ◽

Single Clone

Several human genes related to DNA excision repair (ER) have been isolated via ER cross-species complementation (ERCC) of UV-sensitive CHO cells. We have now isolated and characterized cDNAs for the human ERCC5 gene that complement CHO UV135 cells. The ERCC5 mRNA size is about 4.6 kb. Our available cDNA clones are partial length, and no single clone was active for UV135 complementation. When cDNAs were mixed pairwise with a cosmid clone containing an overlapping 5'-end segment of the ERCC5 gene, DNA transfer produced UV-resistant colonies with 60 to 95% correction of UV resistance relative to either a genomic ERCC5 DNA transformant or the CHO AA8 progenitor cells. cDNA-cosmid transformants regained intermediate levels (20 to 45%) of ER-dependent reactivation of a UV-damaged pSVCATgpt reporter plasmid. Our evidence strongly implicates an in situ recombination mechanism in cDNA-cosmid complementation for ER. The complete deduced amino acid sequence of ERCC5 was reconstructed from several cDNA clones encoding a predicted protein of 1,186 amino acids. The ERCC5 protein has extensive sequence similarities, in bipartite domains A and B, to products of RAD repair genes of two yeasts, Saccharomyces cerevisiae RAD2 and Schizosaccharomyces pombe rad13. Sequence, structural, and functional data taken together indicate that ERCC5 and its relatives are probable functional homologs. A second locus represented by S. cerevisiae YKL510 and S. pombe rad2 genes is structurally distinct from the ERCC5 locus but retains vestigial A and B domain similarities. Our analyses suggest that ERCC5 is a nuclear-localized protein with one or more highly conserved helix-loop-helix segments within domains A and B.

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Ammonia regulation of amino acid permeases in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.3.4.672-683.1983 ◽

1983 ◽

Vol 3 (4) ◽

pp. 672-683

Author(s):

W E Courchesne ◽

B Magasanik

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Cytoplasmic Membrane ◽

Nitrogen Sources ◽

Amino Acid Permease ◽

Inducer Exclusion ◽

Amino Acid Permeases ◽

General Amino Acid Permease ◽

State Growth ◽

And Control

The activities of the proline-specific permease (PUT4) and the general amino acid permease (GAP1) of Saccharomyces cerevisiae vary 70- to 140-fold in response to the nitrogen source of the growth medium. The PUT4 and GAP1 permease activities are regulated by control of synthesis and control of activity. These permeases are irreversibly inactivated by addition of ammonia or glutamine, lowering the activity to that found during steady-state growth on these nitrogen sources. Mutants altered in the regulation of the PUT4 permease (Per-) have been isolated. The mutations in these strains are pleiotropic and affect many other permeases, but have no direct effect on various cytoplasmic enzymes involved in nitrogen assimilation. In strains having one class of mutations (per1), ammonia inactivation of the PUT4 and GAP1 permeases did not occur, whereas glutamate and glutamine inactivation did. Thus, there appear to be two independent inactivation systems, one responding to ammonia and one responding to glutamate (or a metabolite of glutamate). The mutations were found to be nuclear and recessive. The inactivation systems are constitutive and do not require transport of the effector molecules per se, apparently operating on the inside of the cytoplasmic membrane. The ammonia inactivation was found not to require a functional glutamate dehydrogenase (NADP). These mutants were used to show that ammonia exerts control of arginase synthesis largely by inducer exclusion. This may be the primary mode of nitrogen regulation for most nitrogen-regulated enzymes of S. cerevisiae.

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Intracellular Retention of the Corticotrophin-releasing Hormone (CRH) Precursor Within COS-7 Cells

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215549804601012 ◽

1998 ◽

Vol 46 (10) ◽

pp. 1193-1197 ◽

Cited By ~ 1

Author(s):

Marcelo J. Perone ◽

Simon Windeatt ◽

Ewan Morrison ◽

Andy Shering ◽

Peter Tomasec ◽

...

Keyword(s):

Amino Acid ◽

Golgi Apparatus ◽

Amino Acid Sequence ◽

Protein Secretion ◽

Intracellular Trafficking ◽

Secretory Pathway ◽

Intracellular Localization ◽

Primary Amino Acid Sequence ◽

Releasing Hormone ◽

Primary Amino

We investigated the intracellular localization of CRH in transiently transfected COS-7 cells expressing the full-length rat corticotropin-releasing hormone (CRH) precursor cDNA. CRH synthesized by transfected COS-7 cells is mainly stored intracellularly. In contrast, CHO-K1 cells expressing the same CRH precursor stored and released equal amounts of immunoreactive (IR)-CRH. Ultrastructural analysis revealed that CRH is stored in electron-dense aggregates in the RER of transiently transfected COS-7 cells and does not migrate into the Golgi apparatus. On the basis of the different intracellular localization, storage, and release of CRH in COS-7 and CHO-K1 cells, we hypothesize that the intracellular trafficking of CRH within the constitutive secretory pathway for protein secretion not only depends on its primary amino acid sequence but might also be influenced by intracellular conditions or factors.

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